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FINAL YEAR PROJECT 2013
Transcript of FINAL YEAR PROJECT 2013
THE DYNAMICS OF ACETOWHITENING IN RABBIT CORNEAS AND TISSUE-ENGINEERED MODEL OF SQUAMOUS EPITHELIUM
MATERIALS & METHODS
SUPERVISOR: DR STEPHEN MATCHER
FINAL YEAR PROJECT
* Cervical cancer is the eleventh most common cancer in the UK.
* Acetowhitening phenomenon is used in conjunction with Colposcopy for the detection of pre-cancerous lesion.
* What is Acetowhitening effect?
* What is Colposcope?
* Limitation of Colposcopy - sensitive but not specific enough to confirm diagnosis.
* Optical Coherence Tomography (OCT) is an optical signal acquisition method based on low coherence interferometry technique.
* Non-invasive, high spatial resolution and enhanced depth penetration.
* Specific in the detection of micro-structural appearance in biological tissues.
Use OCT to image rabbit cornea and tissue-engineered model of squamous epithelium before and after topical addition of acetic and lactic acid
To produce non-keratinised tissue-engineered model of squamous epithelium.
To characterise the dynamics of acetowhitening effect by quantifying the backscattering intensity at different depths of the multilayer tissues using Image J.
* Tissue-engineered model of squamous epithelium was produced by submerged culturing in Green's Media for 2 weeks.
PROTOCOL FOR OCT IMAGING
Samples were transferred into a well in petri-dish formed by blue tacks.
A reference OCT image was captured before the addition of acids.
Continuous OCT images were then captured upon application of 0.5mL acids in 0.5M, with a frequency of one image every 4 seconds for a total of 240 seconds.
The samples were immediately fixed in 3.7% formaldehyde and processed for immunohistochemistry.
Aim & Objectives
Materials & Methods
Conclusion & Further Work
* Non-inverted and inverted OCT were used in the study for rabbit cornea.
* Non-inverted OCT was used in the study for tissue-engineered skin composites.
REFLECTIVITY STUDY ON MOLECULAR DIFFUSION
VIDEO SHOWING ACETOWHITENING PHENOMENON IN THE FIRST 100 SECONDS
0.5M OF ACETIC ACID
REFLECTIVITY CURVES ACROSS DIFFERENT DEPTHS OF RABBIT CORNEAS
* Why tissue-engineered model of squamous epithelium?
* Why rabbit corneas?
EPITHELIUM WHITENING AND PERMEABILITY OF ACIDS
TISSUE-ENGINEERED SQUAMOUS EPITHELIUM
REFLECTIVITY CURVES ACROSS DIFFERENT DEPTHS OF MULTILAYERS SQUAMOUS EPITHELIUM
* First study on depth-resolved kinetics of acetowhitening.
* Quantitative results indicate a new aspect to the acetowhitening phenomenon.
* Contribute clinically towards the diagnosis of cervical cancer and hence, prevent the need of unnecessary biopsy and introducing long-term monitoring of the disease progression.
 Massad, L.S et al (2009) Journal of Lower Genital Tract Disease 13: 137-144
 Maclean, Allan B (2004) Gynaecologic Oncology 95: 691–694
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 Balas, C et al (2008) Journal of selected topics in quantum electronics 14: 29-42
 Gallwas, Julia K.S et al (2011) Lasers in Surgery and Medicine 43: 206-212
 Kocsis E et al (1991) Journal of Structural Biology 107: 6-14
* Acetowhitening effect could be monitored in a prolonged period of time and across a wider depths.
* For the tissue-engineered skin, a different protocol could be introduced to reduce the artefacts caused by the change in optical path length.
0.5M OF LACTIC ACID
I am truly and sincerely thankful to my project supervisor, Dr Stephen Matcher for providing me tremendous support and guidance to complete the project.
I would like to express my gratitude to Dr Shweta Mittar for her help in culturing the tissue-engineered model of squamous epithelium.
I would like to thank Dr Zeng Hai Lu for facilitating me in using Optical Coherence Tomography throughout the project.